Kidney Week

Abstract: SU-OR03

Incorporation of Urine-Derived Stem Cells into Kidney Organoids Derived from Human Induced Pluripotent Stem Cells

Session Information

• AKI Mechanisms: Research Abstracts
  October 25, 2020 | Location: Simulive
  Abstract Time: 05:00 PM - 07:00 PM

Category: Acute Kidney Injury

• 103 AKI: Mechanisms

Authors

• Bejoy, Julie, Vanderbilt University Medical Center, Nashville, Tennessee, United States

Julie Bejoy, PhD



I am currently working as a research Fellow at Vanderbilt University Medical Center, Nashville,Tennessee. I received my Ph.D. in Biomedical Engineering from Florida A&M University in 2019. During my graduate research I worked on tissue engineering and characterization of the neural cells. I derived neurons and astrocytes from induce pluripotent stem cells and used them for neurological disease modeling, drug screening, and neural tissue regeneration. My research focused on the interactions of different cell signaling pathways in the neural and glial fate of human pluripotent stem cells and corresponding implications for neural organoid formation. During this time I have presented both poster and oral presentations at the Biomedical engineering society (BMES) Conferences, received a best poster award from the Department of Chemical and Biomedical Engineering, FAMU. During my bachelor’s program in India I received a financial grant from the Institution of Engineers (India) for my final year project named “Intestinal Myogram”. We designed an invasive device and studied the effectiveness of the device to measure the intestinal signals. Currently in my post-doctorate research I am applying the skills I achieved during previous years to develop regenerative medicine. I am working on the human urine-derived stem cells, evaluating the possibility of achieving different kidney cell types from these cells using either optimized transfection or using growth factor modulation. I am also working on the generation of IPSC derived kidney organoids which can be used for kidney disease modeling, drug screening, and kidney tissue regeneration.

• Welch, Richard C., Vanderbilt University Medical Center, Nashville, Tennessee, United States

Richard C. Welch,

• Woodard, Lauren Elizabeth, Vanderbilt University Medical Center, Nashville, Tennessee, United States

Lauren Elizabeth Woodard,

Group or Team Name

• Woodard Lab

Background

Donor-derived somatic cells or stem cells can be differentiated into renal cell types for disease modeling, drug screening, or therapeutic studies. Modeling of kidney disease with kidney organoids derived from human induced pluripotent stem cells (hiPSCs) has been termed a “kidney in a dish.” The recent advances in stem cell-based therapies have shown great promise for the treatment of kidney injuries. To evaluate the therapeutic properties, we studied the incorporation of urine-derived stem cells (USCs) into a kidney organoid model of acute kidney injury. UCSs are viable cells from urine which can be expanded in vitro for more than ten passages. There is evidence suggesting that USCs are most likely cultured glomerular parietal epithelial cells.

Methods

For this project, we cultured kidney organoids from fibroblast-derived hiPSCs by the established protocol from Takasato and Little, following optimization.Co-culturing of USCs labeled with a membrane dye and Day 25 kidney organoids revealed that USCs incorporated into the organoids efficiently within two days of the co-culture. For injury models, we established nephrotoxicity in the proximal tubule by adding the nephrotoxic drug Cisplatin (5 μm) at Day 21 of kidney organoid culture.

Results

The kidney organoids derived form IPSCs expressed the kidney cell type markers ECAD (distal tubule), GATA3 (collecting duct), LTL (proximal tubule) and NEPHRIN (Glomeruli) at Day 21.The organoids were then treated with 5 x 10⁴ USCs at Day 22 for 48 hours and evaluated for the expression of kidney injury molecule-1 (KIM-1). Immunostaining revealed that KIM-1 expression was significantly reduced in the organoids treated with USCs compared to the organoids without USCs, suggesting a positive therapeutic impact of USCs. We are currently performing RNAseq on three sets of whole kidney organoids (Control, +Cisplatin, +Cisplatin+USC) to provide detailed interrogation of cellular apoptosis and related signaling pathways in these three different sets of organoids.

Conclusion

The ability of USCs to reduce KIM-1 expression in human kidney organoids suggests that further investigation into the therapeutic potential of USC for treatment of acute kidney injury is warranted.

Funding

• Private Foundation Support